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Regenerative Medicine in Ocular Surface Reconstruction: Advancing Cell-Based Therapies for Limbal Stem Cell Deficiency
de Sousa Cabral, João Victor ; Jirsová, Kateřina (advisor) ; Mahelková, Gabriela (referee) ; Mandys, Václav (referee)
Limbal stem cell deficiency is a disease caused by the impairment of limbal epithelial stem cells (LESCs), leading to the replacement of the corneal surface with nontransparent conjunctiva, and its treatment is not standardly available worldwide. This work focuses on preparing cells for treating both uni- and bilateral forms of the disease using advanced cell- based therapy. It investigates LESCs in various scaffolds (fibrin, nanofibers) and explores non-limbal cell sources like oral mucosal epithelial cells (OMECs) under standard complex and xenobiotic-free culture conditions. The study uses immunofluorescence and gene expression to detect stem cell markers, proliferation, and differentiation capacity of cultured cells. Moreover, long-term OMECs storage with different media and cryoprotective agents and the healing properties of the amniotic membrane (AM) were also examined. We found that adding interleukin-13 to the culture media enhanced LESCs' stemness. LESCs on fibrin gels showed higher expression of stemness markers, while those on polymers expressed more mesenchymal ones. Using standard and xeno-free media, we successfully prepared OMEC-containing cell sheets on fibrin gel substrates. The cultured cells exhibited high expression of stemness genes (ΔNp63α, NGFR, KLF4) and decreased levels of...
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Targeted differenciation of limbal and mesenchymal stem cells and their therapeutic application
Kuthanová, Hana ; Holáň, Vladimír (advisor) ; Indrová, Marie (referee)
The research of stem cells slowly transfers from the experimental to the preclinical and clinical level. They are in the centre of interest thanks to their potential to treat many of severe injuries and genetically determined diseases. However, the clinical application of these cells has to be based on a basic research of their characteristics and differential potential. Adult stem cells are in organism in minor populations in unique niches. In comparison with embryonic and induced pluripotent stem cells, the adult stem cells have lower differential potential but they also tend less to making teratomas. The therapeutic use of differential and transdifferential potential of limbal and mesenchymal stem cells is described here in more detail with focus on their use in damaged ocular surface treatment. Limbal stem cells are the only source of stem cells for corneal epithelium regeneration in most organisms. Deficiency of these stem cells leads to severe eye disorders even to blindness. Nowadays, a transplantation of allogeneic limbal stem cells or allogeneic limbus is the only chance for patients with total limbal stem cell deficiency. In clinical trials with patients with particular limbal stem cell deficiency, autologous limbal stem cells were successfully transplanted. Mesenchymal stem cells derived...
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